Chs chaox



United States Patent 3,123,597 ZO-BIS-(I-IYDROXYMETHYD-ll-QXYGENATED-PREGNANES AND DERIVATIVES THEREOF Daniel Bel-tin, Montrcuge, Seine,Hubert Fritel, Paris,

Georges Muller, Nogent-sur-Marne, Jean Mathien,

Moutiermeii, Robert Joiy, Montmorency, and Jean Jolly,Fontenay-sous-Bois, France, assignors to Reuse]- UCLAF, Paris, France, acorporation of France No Drawing. Filed Mar. 26, 1962, Ser. No. 182,679

Claims priority, application France July 8, 1959 32 Claims. (Cl.260-2395) This invention relates to novel20-bis-(hydroxymethyl)-pregnanes and the process of preparation thereof.The invention more particularly relates to ZO-bis-(hydroxymethyl)-pregnanes selected from the group consistib ECU (III) wherein R isselected from the group consisting of =0 and R is selected from thegroup consisting of :0 and and Ac is selected from the group consistingof hydrogen, an acyl radical of an organic carboxylic acid having 1 to18 carbon atoms and an anion of a mineral acid.

In commonly assigned, copending application Serial No. 88,282, filedFebruary 10, 1961, now Pat. No. 3,052,- 675, it is disclosed thatSa-pregnane compounds which are not oxygenated in the ll-positionpossess cardiotropic activity coupled with a dilation of the coronaries.

The products of the invention are distinguished by their cardiotropicactivity coupled with a dilation action of coronaries. They are usefuleach time that a specific action on the heart muscle is necessary, thisaction being in addition accompanied with a beneficial augmentation ofsanguinine irrigation of the said heart muscle.

It is an object of the invention to produce novel bis- (hydroxymethyl)-1 l-oxylgenated-pregnanes.

It is a further object of theinvention to provide novel intermediatesfor the preparation of 20-bis-(hydroxymethyl) -1 l-oxygenated-pregnanes;

Itis another object of the invention to provide novel processes for thepreparation of 20-bis-(hydroxymethyl)- 1 l-oxygenated-pregnanes.

"It is an additional object of the invention to prepare pharmaceuticalcompositions comprising 20-bis-(hydroxymethyD-ll-oxygenated-pregnanesfor the treatment of heart conditions.

The products of the invention are prepared starting from3a-acetoxy-ZO-formyl-Sfl-pregnane l1 one which has a melting point of190 C. The said compound unr dergoes the Tollerrs condensation withformol in the pres-v maldehyde and potassium hydroxide at roomtemperature and recovering the desired product.

Upon reduction of 20-bis- (hydroxymethyl)-5fl-pregn-anc-3oa-Ul-11-One inan inert organic solvent, ZO-bis-(hydroxymethyl)-B-pregnane-3a,l1;8-dio1 is formed. Suitable reducing agents for thereduction are lithium aluminum hydride and alkali metal borohydridessuch as potassium borohydride, sodium borohydride and lithiumborohydride. An example of a suitable inert solvent for the reaction istetrahydrofiuran. The reaction scheme is outlined in Table I.

TABLE I CH3" CHQOH o'Ha CHqOH fIU CHmH The process for the preparationof 20-bis-(acy1oxymethyl)s-5 8-pregnane-3,1l-diones comprises reacting20- bis-(hydroxymethyl) 5B -pregnane-3u-ol-11-one with a ketone oraldehyde to form the al-kylidene or larylalkylidone of 20 bis(hydroxymethyl)-5fl-pregnane-3mo -11- one, oxidizing the latter in the 3hydroxy position to form the corresponding alkylidene or arylalkylideneof 20-bis- (hydroxymethyl)-5,B pregnane -'3 ,11 dione, hydrolyzing saidcompound underacidic conditions to form ZO-bis(hydroxyrnethyl)-5,8-pregnane 3,1l-dione. The free alcohol may then beacylated with amineral or organic acid to. form the corresponding20'-bis-(-acyl'oxymethyl)- 5,8-pregnane-3,1 l-dione.

A preferred process for the preparation of 20-bis- (acyloxymethyl)5fi-pregnane-3,11=diones comprises reactingZO-bis-(hydroxymethyl)-55-pregnane-3u-ol-1l-one with acetone at roomtemperature in the presence of perchlon'c acid to form the acetonide ofZO-bis-(hydroxymethyl)-5B-pregnane-3a-o1-1l one, oxidizing saidacetonideat a low temperature ofthe order of to 10 C. with; sulfochromicacid to form the acetonide of 20bis (hydroxymethyD-Sfi-pregnane 3,11dio'ne, reacting the latteriwith. a refluxing solution of sulfuric acidin ethanol to form 20-bis(hydroxymethyl)-5B-pregnane 3,1l-dione whichcan bereacted with fuming 'nitnic acid attemperatures of -0 to -15 9 C.-to formZO-bis-(nitratomethyl)+5 6- pregnane-3,11-dione. Other acids maybe usedfor the acylationt The reaction scheme is shown in Table 'II.

7 TABLE II H C. g H

CH3 CHzOH o f onion CH3 CHZOAC flj OHzOAc wherein R and R are selectedfrom the group consisting of hydrogen,. phenyl, phenyl-substituted loweralkyl and lower alkyl and Ac has the above definition.

Other ZO-bis-(acyioxymethyl)ddpregnanes of the invention may be producedby preparing alkylidene 0r arylalkylidene of 20-bis-(hydroxymethyl) 5,8pregnane- 301-01-11-0116 as described above, acylating the latter withan acylating agent such as an acid anhydride in an inert solvent to formthe alkyliderie or arylalkylidene of 3aacyloxy 20 bis(hydroxymethyl)-5fi-pregnane-1l-one, hydrolyzing said product underacidic conditions to form 3zx-acyloxy-20-bis (hydroxymethyl) 5Bpregnane-l 1- one, acylating the latter to form3a-acyloxy-20-bis-(acyloXymethyl)-5,8-pregnane-1l-one, and saponifyingthe said compound to form ZO-bis-(acyloxymethyl)-5/3-pregnane-3a-ol-11-one. This compound may be acylated with a mineral acid or anorganic acid to form another 30:-acyloxy-ZO-bis-(acyioxymethyl)-5fi-pregnane-1l-one or it may be reducedto form 20bis-(acyloxymethyl)-5p-pregnane-3 06,11fidi01.

A preferred process for the preparation of these compounds comprisespreparing the acetonide of ZO-bis-(hydroxymethyl) 5,8 pregnane 3aol-ll-one as described above, acetylating said acetonide with aceticacid anhydride in pyridine to form the acetonide of 3ot-acetoxy-20-bis-(hydroxymethyl) 5B pregnane-l l-one, reacting said product withaqueous acetic acid at room temperature to form 3 a-acetoxyifl-bis-(hydroxymethyl -5fi-pregnane-1 1- one, reacting the latter with fumingnitric acid at temperatures between 5 and 15 C. to form 3a-acetoxy-ZO-bis-(nitratomethyl)-5,B-pregnane-1l-one and saponifying to form20bis-(nitratomethyl)-5fi-pregnane-3a-ol-11- one.

ZO-bis-(nitratomethyl) Sfl-pregnane-Iia-ol-ll-one may be acylated withfuming nitric acid at low temperatures: to 1 form3a-nitr21t0-20-bis-(nitratomethyl) 5B pregnane-- ll-one or may beacylated with sulfuric acid anhydrideor sulfuric chlorohydrin inpyridine to form the pyridine salt of3a-sulfato-20-bis-(nitratomethyl)-5/3-pregnane-l1- one which can beconverted by double decomposition into its alkali metal salt.

The outline of the prepartion of these compounds is. illustrated inTable III.

CH3 CHzOH onion CH: GH1O\ Ra omo \R4 RO-- i7 CH3 CHzOH fl \OH2OH no" 3 n0H3 CHzOAc CHzOAc CH3 CHnOAQ AeO-- wherein R R and Ac have the abovedefinitions and R is the acyl radical of an organic carboxylic acidhaving 1 to 18 carbon atoms.

The unsaturated compounds of Formula III are produced by forming thealkylidene or arylalkylidene of 20- bis-(hydroxymethyl) 11oxygenated-Sfi-pregnane-I:-one as described previously, reacting thelatter under acidic conditions to form 20-bis-(hydroxymethy1) 11oxygenated-5fi-pregnane-3-one, acylating said compound with an organicacid to form 20-bis-(acyloXymethyl)-1l-oxygenated-Sfl-pregnane-S-one,brominating the said product to form4-bromo-20-bis-(acyloxymethyl)-1I-oXygenated-SB- pregnane-3-one,dehydrobrominating the latter to form 20-bis (acyloxymethyl) 11oxygenated-A -pregnene-3- one, saponifying the unsaturated compound toform 20- bis-(hydroxymethyl) 11 oXygenated-A -pregnene-3-one which maybe acylated with an organic carboxylic acid having 1 to 18 carbon atomsor with a mineral acid to form 20-bis-(acyloxymethyl) -1 1-oXygenated-A-pregnene- 3-one.

A preferred process for the preparation of the compounds of Formula HIcomprises forming the acetonide ofZO-bis-(hydroxymethyl)-5B-pregnane-3,1l-dione as decribed previously,refluxing the latter in ethanolic solution of sulfuric acid to form20-bis-(hydroxymethyD-SB- pregnane-3,l1-dione, acylating said compoundwith acetic acid to form ZO-bis-(acetoxymethyl)-5fipregnane-3,1ldione,brominating the latter with a solution of bromine in acetic acid attemperatures about 50 to C. to form 4bromo-20-bis-(acetoxymethyl)-5,8-pregnane-3,11- dione,dehydrobrominating the said product with a mixture of lithium bromideand lithium carbonate in a di-lower alkyl carboxamide such asdimethylformamide to form ZO-bis-(acetoxymethyl)-A-pregr1ene-3,11-dione, saponifying the latter with alkali metalhydroxide at room temperature to form 20bis-(hydroxymethyl)-Apregnene-3,1l-dione which may be acylated by reacting with fuming nitricacid at temperatures about --5 and '15 C. to formZO-bisJ-(nitratomethyl)-A -pregnene- I.g/ll-dione. This reaction schemeis illustrated by Table 7 TABLE IV CH3 cnzon RZfUX CH3 onion R2:( 'UCH2OR Ca CH2OR onion E H Br CH5 omon mfIUX q/ CH: OHQOH R2=(\U CHZOH o:I

CH; CHaOAc RZ{\IU OH2OAG 0: i

wherein R, R and Ac have the above definitions.

The unsaturated compounds of Formula II may be prepared by formingZO-bis-(acyloxymethyD-ll-oxygenated-Sfi-pregnane-Ia-one as describedpreviously, brominating the latter to form2,4-dibromo--bis(acyloxymethyl) 11- oXygenated-5fi-pregnane-3-one,.dehydrobrominating the latter to form ZO-bis-(acyloxymethyl)-l'1-oXygenated-A -pregnadiene-3-one, saponifying the latter to form20-bis-(hydroxymethyl)-1l-oxygenated-A pregnadiene-3-one which may beacylated with an organic 8 carboxylic acid having 1 to 18 carbon atomsor a mineral acid to form 20-bis-(acyloxymethyl)-11-0xygenated-Apregnadiene-3-one.

A preferred process for the preparation of the unsaturated compounds ofFormula II comprises forming 20-bis- (acetoxymethyl) -ll-0Xygenated-5/3-pregnane-3 -one as previously described, brominatingthe latter with a solution of bromine in acetic acid to form2,4-dibromo-20-bis- (acetoxymethyl) -1 l-oxygenated -5B-pregnane-3-one,dehydrobrominating said product with a mixture of lithium bromide andlithium carbonate in a solvent such as dimethylformamide to formZO-bis-(acetoxymethyD-l1- oxygenated-A -pregnadiene-3-one, saponifyingthe latter with an alkaline solution to from ZO-bis-(hyroxymethyD-11-oxygenated-A -pregnadiene-3-one which may be further acylated on acidsuch as fuming nitric acid to form the corresponding acylate such asZO-bis-(nitratomethyl)-1l-oxygenated-A -pregnadiene-3-one. Table Villustrates the method of preparation.

TABLE V CH OR C53 GHzOR R I/\IU/ Z Br g H Br CH3 CHzOR crnon /\{3/ J CH3CHgOH CH3 CHzOAc wherein R, R and Ac have the above-definitions.

CH3 CH O CH C f'j omo om CH; CH OH NazOaPO to form 30:dibenzylphosphato-ZO-bis-(hydroxymethyl)- 5B-pregnane-ll-one, subjectingthe latter to hydrogenolysis in the presence of a palladium catalyst toform 30:- phosphato-ZO-bis-(hydroxymethyl) 5 ,8 pregnane-ll-one whichcan then be changed to its alkali metal salt or esterified with anorganic carboxylic acid having 1 to 18 carbon atoms or a mineral acid.

A preferred process for the preparation of 3a-phosphato-20-bis-(hydroxymethyl)-5/3-pregnane-l l-one comprises reacting the acetonide of20-bis-(hydroxymethyl)- 5 8-pregnane-3u-ol-11-one with dibenzylchlorophosphonate in an ether solution to form the acetonide of3oc-dibenzylphosphato 20 bis (hydroxymethyD-SB-pregnane-ll-one,hydrolyzing the latter with hydrochloric acid in an aqueous alcoholsolvent to form3a-dibenzylphosphato-ZO-bis-(hydroxymethyl)-5,8-pregnane-l l-one,subjecting the latter to hydrogenolysis in the presence of palladizedcarbon black in methanol to form SOL-PhOSPhfllO-20-bis-(hydroxymethyl)-5fi-pregnane-l1-one. If the latter is reactedwith sodium methanolate in methanol the disodium salt of3a-phosphato-20-rbis-(hydroxymethyl)- S/S-pregnane-ll-one is formed. Ifinstead the said phosphato compound is reacted with an esterifying agentsuch as fuming nitric acid, the corresponding ester such as 300phosphato-ZO-bis-(nitratomethyl)-5fi-pregnane-1l- 10 one is formed. Thereaction is illustrated in Table VI. The alkali metal salts of3a-phosphato-20-bis-(hydroxymethy1)-5B-pregnane11-one are particularlyadapted for administration by transcutaneous injection due to theirwater solubility.

TABLE VI (Co aC 2)zOz CH CHgOH CHzOH C 3 CHaONOg fU CaONO.

wherein R and R have the above definitions.

The terms Ac and acyl used hereinbefore may represent the acyl radicalof an organic carboxylic acid having 1 to 18 carbon atoms or a mineralacid. Suitable organic carboxylic acids are alkanoic or alkenoic acidssuch as acetic acid, trimethylacetic acid, propionic acid, 4,4-dimethy1pentanoic acid, lo-undecenoic acid, cycloalkyl alkanoic acids such asfi-cyclopentyl propionic acid; arylalkanoic acids such as phenylpropionic acid; cycloalkanoic acids such as hexahydrobenzoic acid andhexahydroterephthalic acid; and phenyl carboxylic acids such as benzoicacid and 3,5-dinitrobenzoic acid. Examples of suitable mineral acids arenitric acid, phosphoric acid and sulfuric acid.

The ketone or aldehyde reactant may be aliphatic or aromatic. Examplesof suitable ketones are acetone, methylethyl ketone, ethylpropyl ketoneand acetophenone. Examples of suitable aldehydes are formaldehyde,acetaldehyde, 'butynaldehyde, benzaldehyde and phenylacetaldehyde. Whenthe aldehyde is aromatic such as benzaldehyde, the phenyl orphenylalkylidene of the ZO-bis-(hydroxymethyl)-pregnanes can betransformed into the free ZO-bis-(hydroxymethyl)-preg11anes by simplehydrogen- 11 embodiments to illustrate'the invention.

be limited to the specific embodiments.

Example I.Preparalin of ZO-Bis-(Hydroxymethyl)*-fi-Pregnane-3oc-Ol-11-One 3 gm. of-formyl-3zx-acetoxy-5fi-pregnane-1l-one, melting point: 190 C., wereintroduced into a mixture of 30 cc. of methanol, 3 cc. of a 30% aqueoussolution of formaldehyde and 1.5 cc. of a 50% potassium hydroxidesolution. The mixture was stirred at C. until solution was complete,then allowed to stand for sixteen hours at a temperature of 25 C.Thereafter, 100 cc. of water were added and the mixture was vacuumfiltered. The filter cake was washed with water and dried in an oven at100 C. 2.74 gm. (94%) of20-bis-(hydroxymethyl)-5fi-pregnane-3x-ol-1l-one, having a melting pointof 209 C. and a specific rotation [a] =+48i5 (c.-=0.5% in ethanol) wereobtained.

The product, which is new, is found in the form of small colorlessneedles. It is insoluble in water, very slightly solublein ether,benzene and chloroform, and soluble in alcohol.

AnaZysis.C H O molecular weight=378.54. Calculated: C, 72.97%; H,10.12%. Found: C,73.1%; H, 9.9%.

Example II.Prepzzrati0n of 20-Bz's- N itratomethyl -5 ,8-Pregnane-3a-Ol-l 1 -One STEP A: PREPARATION OF ACETONIDE OF ZO-BIS-(HYDROXYMETHYL) -5fl-PREGNANE-3aOL-1l-ONE 8 gm. ofZO-bis-(hydroxymethyl)-5{3-pregnane-3a-ol- 11-one were placed insuspension in 400 cc. of acetone, agitated at room temperature and 1.2cc. of 65% perchloric acid was added. After complete solution of thesteroid compound, the reaction mixture was allowed to stand at roomtemperature for two hours. Then.1.6

gm. of sodium bicarbonate and 800 cc. of ice water.

were added. The precipitate obtained was vacuum filtered at 0 C., washedwith water and dried at 100 C.

The raw product was dissolved at reflux in ethanol containing 2% ofpyridine. The solution was then treated with animal charcoal, filtered,concentrated and cooled for one hour at 0 C. The crystals obtained werewashed with iced ethanol, then dried at 100 C. 5.8 gm. of the acetonideof 20-bis-'(hydroxymethyl)-5B-pregnane-3u-ol-11-one were obtained, beinga yield of 66%. The product had a melting point of 206.5" C. and aspecific rotation of [a] ==-|-30 C. (c.=1% in chloroform).

The product was soluble in acetone, benzene, chloroform and ethanol,insoluble in'water and ether.

Analysis.C H O molecular weight=4l8.60. Calculated: C, 74.60%; H,10.11%. Found: C, 74.3%; H, 10%.

This compound is not described in the literature.

The starting compound was prepared by the condensation of formol with20-formyl-3a-acetoXy-SfI-pregnanell-one according to the Tollensreaction in the presence of potassium hydroxide.

STEP B: PREPARATION OF 3a-ACETOXY-20-BIS-(HY- DROXYMETHYL)5fi-PREGNANE-l1-ONE 5 gm. of the compound prepared in Step A wereintroduced into 30 cc. of pyridine and the solution was made lukewarm toeffect its dissolution. The solution was then cooled to room temperatureand 10 cc. of acetic anhydride were added.

The reaction mixture was allowed to remain for two hours at roomtemperature and was then poured into 300 cc. of slightly alkaline icewater (pH 8). The gummy precipitate obtained was extracted three timeswith 60 cc. aliquots of ether. The ethereal extracts were united, washedwith water until the wash water was neutral, then dried over magnesiumsulfate, filteredand evapo- However, it should be understood that theinventionis not intended to 12 e ratedto dryness under vacuum. A brownresin was obtained with a theoretical yield.

The resin obtained was introduced in 165 cc. of 60% acetic acid. Thereaction mixture was agitated-for-thirtyminutes at room temperature,then for one hour at 65- C. and the mixture was then cooled at roomtemperature. 300- mg. of animal charcoal were added, the mixturefiltered and the filtrate washed with 60% acetic acid.

The combined filtrates were poured into 750 cc. of iced Water. The gummyresidue formed was extracted by ethyl acetate. The organic phase wasWashed successively with water, then with a 0.10 N sodium hydroxidesolution and again with water, then dried over sodium sulfate andevaporated under vacuum.

A gummy residue was obtained which crystallized by titration withisopropyl ether. The crystalline precipitate was vacuum filtered at 0C., washed with isopropyl ether and dried at 60 C.

Recrystallization was effected from..-carbon tetrachloride. 2.6 gm. of3m-acetoxy-20-bis-(hydroxyrnethyl)5,8- pregnane-ll-one were obtained,being a yield of 53% and had a melting point of 211 C. and a specificrotation [a] =+61.7 (c.:l% in chloroform).

The product was soluble in acetone, benzene, chloroform, ethanol, ether,slightly soluble in carbon tetrachloride, insoluble in water and diluteaqueous acids and alkalis.

Awalysis.C H O molecular weight=420.57. Calculated: C, 71.39%; H, 9.50%.Found: C, 71.2%; H, 9.4%.

This compound is not described in the literature.

STEP 0; PREPARATION OF 3a-ACETOXY-20-BIS-(NITRATOMETHYL)-5fl-PREGNANE-11-ONE 3.2 cc. of 48 B. nitric acid wereintroduced slowly into 11.5 cc. of acetic anhydride cooled to 10 C. Thenover a period of five minutes under agitation and under nitrogen at 10C., a solution of 1 gm. of30aacetoxy-20-bis-(hydroxymethyl)-5,6-pregnane-1l-one in 6 cc. ofchloroform was added. The agitation of the reaction mixture wascontinued for twenty minutes at a temperature'between about 5 and 10 C.and then the mixture was'poured under agitation in cc. of iced water.

The aqueous phase was extracted with methylene chloride. The organicphase was washed successively with water, with a solution of 2% ofsodium bicarbonate and again with water until the wash waters wereneutral. The solution was filtered, dried over sodium sulfate andevaporated under vacuum. The gummy residue crystallized by addition ofseveral drops of ethanol. It was homogenized by the addition ofisopropyl ether and heating until boiling. The suspension was cooled,the crys-- STEP D: PREPARATION OF 20BIS-(NITRATOMETH=YL)-5flPREGNANE-3a-OL-11-ONE 1.27 gm. of the compound prepared in Step Cabove were introduced into 50 cc. of ethanol, 3.7 cc. of water and 0.75cc. of a 10 N sodium hydroxide solution. The reaction mixture wasagitated under nitrogen at room temperature for three and one-halfhours. The reaction mixture was then poured into 250 cc. of iced water.The raw product was extracted three times with 40 cc. aliquots of etherafter addition of 20 cc. of a saturated sodium chloride solution.

The ethereal solution was washed with water, dried over sodium sulfate,filtered and evaporated under vacuum. The residue was dissolved in 2 cc.of toluene at 70 C., then cooled to room temperature in order to causecrystallization. The crystals obtained were vacuum filtered, washed withtoluene and dried at 60 C. 1.21 gm. of 20- bis(nitratomethyl)5,B-pregnane-3a-ol-1l-one solvated with a half moleculeof toluene were obtained, being a yield of 96% The solvate had a meltingpoint of 118 C. and a specific rotation [a] =l-15.5 C. (c.=1% indioxane).

The product was soluble in ethanol, ether, acetone and chloroform,slightly soluble in benzene, insoluble in water and dilute aqueous acidsand alkalis.

Analysis.-C H O N +%C H molecular weight: 514.60. Calculated: C, 61.85%;H, 7.83%; N, 5.44%. Found: C, 61.9%; H, 7.8%; N, 5.1%.

Toluene was eliminated in the following fashion: The solvate wasdissolved in ethanol, the alcohol was distilled and ethanol was addedanew. The solution was then con centrated, water added and allowed toremain overnight at C. The precipitate was then vacuum filtered anddried at 60 C.

This compound is not described in the literature.

Example III.Preparation of ZO-Bis-(Hydroxymethyl) 5,8-Pregnane-3u,1 1B-D iol Into 30 cc. of tetrahydrofuran there were introduced 800 mg. oflithium aluminum hydride while maintaining the temperature at 18 C. by awater-ice bath. Then, over fifteen minutes at the same temperature, thefollowing solution was added:

50 cc. of tetrahydrofuran 800 mg. of 20-bis-(hydroxymethyl)-5fi-pregnane-3 a-ol-l 1- one Then another 15 cc. of tetrahydrofuranwere added and the reaction mixture was agitated for four hours at roomtemperature under nitrogen. The excess lithium aluminum hydride wasdestroyed by the addition of 16 cc. of ethyl acetate. About 18 cc. of asaturated solution of sodium chloride diluted in half was thenintroduced under agitation. The mineral precipitate formed was separatedfrom the organic phase, then extracted with ethyl acetate. The organicsolutions were combined and then washed with a saturated solution ofsodium chloride diluted in half, dried over sodium sulfate andevaporated under vacuum.

The resin obtained was dissolved in 48 cc. of ethanol. The solution wasfiltered and concentrated to about 8 cc. and the resin was precipitatedfrom the hot solution by the addition of 10 cc. of water. Theprecipitate was then vacuum filtered, washed with iced 50% alcohol anddried at 80 C.

The crystals obtained were dissolved in a mixture of toluene andalcohol. The solution obtained was concentrated and the productcrystallized from solution by cooling. 374 mg. ofZO-bis-(hydroxymethyl)-5[3-pregnane-3 a, 11,6-diol were obtained, beinga yield of 48%. The said product had a melting point of 210 C. and aspecific rotation [a] =+35.8 (c.=1% in ethanol).

By recovery of a second batch of crystals starting from the motherliquors of the alcohol purification, the yield was increased by 13 Thisgave a total yield of 61% The product was soluble in alcohol, slightlysoluble in toluene and acetone, insoluble in water, ether, benzene andchloroform.

Analysis.-C H O molecular weight=380.55. Calculated: C, 72.59%; H,10.59%. Found: C, 72.9%; H, 10.6%.

The compound is not described in the literature.

The starting compound was prepared by the condensation of forrnyl in theTollens reaction on 20-formyl-3a- 14' acetoxy-Sfl-pregnane-ll-one in thepresence of potassium hydroxide.

Example lV.Preparati0n of the 3-Potassium Sulfate ofZO-Bis-(Nitratomethyl) 5,B-Pregnane-3ot-Ol-1I-One 500 mg. of20-bis-(nitratomethy1)-5[3-pregnane-3a-olll-one were dissolved in 5 cc.of chloroform and 2.5 cc. of anhydrous pyridine. Then, under agitationand under nitrogen at an interior temperature between 0 and +5 C., 14cc. of a 1% solution of sulfuric chlorohydrin in chloroform were added.The reaction mixture was allowed to remain overnight under agitation andunder nitrogen at room temperature. The reaction mixture was filteredand the precipitate was washed with chloroform. The filtrates were thenreunited and 50 cc. of ether added thereto.

The gummy precipitate was separated, dried under vacuum, then taken upwith 50 cc. of ether containing 2% ethanol and triturated until theappearance of crystals. The mixture was vacuum filtered and then washedseveral times with water. 545 mg. of the pyridene salt of the 3-sulfateof 20-bis-(nitratomethyl)-5fi-pregnane-3a-ol-11- one were obtainedhaving a melting point of about 160 C. upon recrystallization fromethanol.

The product obtained was dissolved in 65 cc. of ethanol and the solutionwas concentrated under vacuum in the presence of nitrogen and at roomtemperature until it reached a volume of about 10 cc. 0.75 cc. of asolution containing 11.4% potassium acetate in ethanol were then added.The reaction mixture was maintained for two hours under agitation at atemperature on the order of 0 C. Then it was vacuum filtered. The filtercake was washed by trituration with iced ethanol and anhydrous ether andthe crystals were dried. 390 mg. of 3-potassium sulfate ofQO-bis-(nitratomethyl)-5B-pregnane-3a-ol-l1- one were obtained having aspecific rotation (c.=1% in water).

This compound was soluble in water, acetone, chloroform, ethanol andisopropanol, insoluble in ether and benzene.

Analysis.--C H O N SK; molecular weight=586.7. Calculated: C, 47.08%; H,6.01%; K, 6.66%. Found; C, 47.1%;H, 6.0%;K, 6.5%.

This compound is not described in the literature.

The starting compound was prepared by blocking the hydroxyls of themethylenes of 20-bis-(hydroxymethyl)- 5p-pregnane-3a-ol-1l-one, followedby acetoxylation in the 3-position, deblockage of the primary hydroxylgroups, and finally nitration of these.

Example V.-Preparati0n of 3 a-N z'trato-ZO-Bis- (N itratomethyl) -53-Pregnane-1 1 -One 500 mg. of20-bis-(hydroxymethyl)-5,8-pregnane-3a-ol- 11-one were introduced into10 cc. of acetic anhydride and 2.6 cc. of fuming nitric acid cooled to10 C. The mixture was agitated for twenty-five minutes at a temperaturebetween about -5 and 10 C. and then the reaction mixture was poured intocc. of iced water. The precipitate formed was agitated then for an hourat a temperature between about 0 and +5 C. The mixture was vac uumfiltered and the filter cake was washed with a solution of 10% sodiumdicarbonate, then with water until the wash Waters were neutral, anddried under vacuum.

The raw product was purified by dissolution in acetic acid andprecipitation by addition of water, then vacuum filtering, washing anddrying, as has already been described above. 485 mg. of3a-nitrato-ZO-bis-(nitratomethyl)-5;8-pregnane-11-one were obtained. Afurther purification was eifected by chromatography on silica gel andelution with methylene chloride containing 8-10% of carbontetrachloride, then by dissolution in acetone or methanol, concentrationof the filtered solution, addition of petroleum ether andcrystallization.

A product was obtained whose melting point was .100-

15 105 C. and whose specific rotation was [a] =+42 i3 (c.=1% indioxane). It was soluble in acetone, benzene, chloroform, acetic acidand methylene chloride, soluble in the hot in alcohol, insoluble inwater.

Analysis. -C H O N molecular weight=513.53. Calculated: C, 53.8%; H,6.87%; N, 8.18%. Found: C, 54.9%; H, 6.8%; N, 7.5%.

This compound is not described in the literature.

This starting compound was prepared by a condensation of forrnol on20-formyl-3a-acetoxy-SB-pregnane-llone by Tollens reaction in thepresence of potassium hydr'oxide.

Example VI.Preparatin of ZO-Bis-(Nitratomethyl)55- Pregnane-3,11 dioneSTEP A: PREPARIION OF THE ACETONIDE 01? 20-1318-(HYDROXYMETHYL)=513-PREGNANE-3,11-DIONE 3gm. of the acetonide of20-bis-(hydroxymethyl)-5fipregnane-3 a-ol-1l-one were dissolved in 600cc. of pure acetone and the solution obtained was cooled until itreached a temperature between about 0 and +5 C. Then 1. 85 cc. of thefollowing sulfochromic solution was added under agitation and undernitrogen:

Chromic acid gm 13.36 Concentrated sulfuric acid cc 11.5 Watersuificient to make cc 50 The addition was made in three steps, withintervals of five minutes between each step.

The agitation was continued at the same temperature for ten minutes. 6cc. of methanol and 6 gm. of dried sodium carbonate were added to thereaction mixture. The agitation was continued for another twenty minutesand the mixture was filtered. The residue was Washed with acetone. Thefiltrate was decolorized by additon of 30 gm. of alumina under agitationfor fifteen minutes, filtered again, 1 cc. of pyridine added to thefiltrate and the solvents removed under vacuum. A resinous residue wasobtained.

The resin was then dissolved in 7 cc. of methanol and the solution wascooled with ice for one hour. The crystals obtained were vacuumfiltered, washed'with isopropyl ether and .dried at60 'C. 1.145 gm. ofthe acetonide of 20-bis-(hydroxymethyl)SB-pregnane-ll1- dione wereobtained. By working up of the product from the mother liquor, a secondbatch of the product was recovered with a weight of 0.59 gm. being atotal of 1.735

The raw product was recrystallized from isopropyl ether and had amelting point of 175. C. and a specific rotation [a] =-}36.5 (c.=l% indioxane).

The product was present in the form of colorless crystals. It wassoluble in alcohol, ether, acetone, benzene andchloroform, insoluble inwater and in dilute aqueous alkalis. It was decomposed in dilute aqueousacids.

Analysis'.C H O molecular weight=4l6.5 8. Calculated: C, 74.96%; H,9.68%. Found: C, 74.9%; H, 9;7%.

This compound is not described in the literature.

The starting material was prepared by blocking the hydroxyls of themethylenes in the 20-position of 20-bis- (hydroxymethyl) -5,8-pregnane-3oc-Ol-l l-one.

STEP B: PREPARATION OF 2O -BIS(HYDROXY- METHYL) -5fl-PREGNANE-3,11-DIONE2 gm. of the acetonide of 20-bis-(hydroxymethyD- 5,8-pregnane-3,ll-dionewere added under agitation to a solution of 1.6 cc. of normal sulfuricacid in 80 cc. of 50% ethanol. The reaction mixture was then carried toreflux for a period of about thirty minutes. Thereafter, it wasneutralized with 290 mg. of sodium bicarbonate. 35 cc. of the alcoholwas removed by distillation under vacuum and under agitation. Thereafter30 .0 Of Water were added and the distillation contin'u'ed. Oily dropswere formed which were transformed into a gum and thereafter set up in awhite precipitate. Another 20 cc. of water were then added and themixture agitated for forty-five minutes at room temperature. Thereaction mixture was then iced, vacuum filtered and the precipitatewashed with water and dried at 60 C. 1.8 gm. ofZO-bis-(hydroxymethyl)-5fl-pregnane-3,l1- dione were obtained.

The raw product was purified by solution at reflux in methanol,precipitation from warm solution by water and successive extractionswith isopropyl ether at reflux temperature. This product had a meltingpoint of 169 C. and a specific rotation [a] =]48.2i3 (c.-=l% indioxane). It was present in the form of colorless crystals which weresoluble in ethanol, acetone, benzene, chloroform and toluene, insolublein water, ether and dilute aqueous acids and alkalis.

Analysis.C I-I O molecular weight=376.52. Calculated: C, 73.36%; H,9.64%. Found: C, 73.2%; H, 9.6%.

This compound is not described in the literature.

STEP C: PREPARATION OF ZO-BIS-(NITRATOMETHYL)- 'Efi-PREGNANE-li,11-DIONE1.6 cc. of 48 B. nitric acid were introduced slowly into 6 cc. of aceticanhydride cooled to -l0 C. T hereafter, a solution of 500 mg. of20-bis-(hydroxymethyl)- 5fi-pregnane-3,1l-dione in 5 cc. of chloroformand 0.25 cc. of'acetic acid were added slowly under agitation and underan atmosphere of nitrogen at a temperature between about 5 and l0 C. Thereaction mixture was maintained under agitation and under nitrogen for aperiod of thirty minutes at a temperature between about 5 and 10 C. Thenit was poured, always under agitation, into 60 cc. of iced water.

The aqueous layer was extracted with three 15 cc. aliquots of methylenechloride. The organic phases were combined, washed successively withwater then with a 2% sodium bicarbonate solution, and again with water,until the wash waters were neutral. The organic solution was dried oversodium, sulfate, filtered and evaporated to dryness. 540 mg. ofZO-bis-(nitratomethyl)Sfipregnane-3,l1-dione were obtained, being ayield of 87%.

The raw product was purified by chromatography on silica gel withelution with methylene chloride, then recrystallized from carbontetrachloride or isopropanol. The purified product had a melting pointof 185-187" C. and a specific rotation [a] =|-17i3 (c.=1% in dioxane).

The product was present in the form of colorless crystals, soluble inacetone, benzene and chloroform, slightly soluble in alcohol, insolublein water and diluteaqueous acids.

Analysis.C H O N molecular weight=466.52. Calculated: C, 59.21%; H,7.35%; N, 6.01%. Found: C, 59.2%; H, 7.3%; N, 5.9%.

This compound is not described in the literature.

Example VII.Preparati0n of the ZO-Bz's-(Nitratomethyl) -A -Pregnene-3, 1l-Dione STEP A: PREPARATION OF THE 20-BIS-(ACETOXY- METHYL)ifi-PREGNANE-Iill-DIONE 1.35 gm. of ZO-bis-(hydroxymethyl)-58-pregnane-3,1ldione were introduced into 8.1 cc. of pyridine and 2.7cc. of acetic anhydride. The reaction mixture was maintained for aperiod of two hours at room temperature and then poured into cc. of icedwater. The gummy residue formed was extracted several times by benzene.The extracts were combined, then washed successively with a normalsolution of hydrochloric acid, with water, with a N/lO normal solutionof sodium hydroxide and finally with water, until the wash waters wereneutral.

The organic phase was dried over sodium sulfate, filtered and evaporatedto dryness under vacuum. The residue was taken up in ethanol andcrystallized. 1.47 gm. of ZO-bis-(acetoxymethyl)-5,6-pregnane-3,ll-dionewere obtained which was recrystallized in aqueous ethanol. Thepurification was effected by a new recrystallization in the samesolvent. The product had a melting point of 135 C. and a specificrotation [a] ='}-25.3 (c.=1% in dioxane).

The product was soluble in acetone, benzene and chloroform, soluble inthe hot in ethanol, insoluble in water and ether.

Analysis.C H O molecular weight=460.59. Calculated: C, 70.40%; H, 8.75%.Found: C, 70.6%; H, 8.8%.

This compound is not described in the literature.

The starting compound was prepared by deblocking of the hydroxyls of themethylenes in the 20-position of the acetonide of20-bis(hydroxymethyl)-5fl-pregnane-3,11- dione.

STEP B: PREPARATION OF THE 20-BIS-(ACETOXY- METHYL) N-PREGNENE-3,11'D'ION\E 1.578 gm. of 20-bis-(acetoxymethyl)-5fi-pregnane-3,11-dione were introduced into 9.5 cc. of acetic acid and heated to 60 C12in order to obtain a dissolution. Bromination was started by addition oftwo drops of hydrobromic acid, then introduction of 6.7 cc. of thefollowing solution:

Pure bromine 1.5 Acetic acid 48.5

The bromination required thirty-five to forty minutes approximately andwas eifected at a temperature of 60i2. The heating was stopped and aftercooling 15 cc. of water were added and the solution was allowed toremain under agitation overnight at room temperature.

The gummy residue was vacuum filtered, washed with water until the washwater was neutral and dried under vacuum. Raw4-bromo-20-bis-(-acetoxyrnethyl)Sfipregnane-3,11-dione was obtained,which was used as such for the following step of the synthesis. Itsmelting point was between 80 and 100 (3.; percent bromine=15.3.

1.093 gm. of lithium bromide were introduced into 12.8 cc. ofdimethyl-formamide. The solution was heated under agitation and undernitrogen to 80 C. and there is then added:

Lithium carbonate 0.890 Above raw diacet-ate 1.780

Cc. Pure anhydrous pyridine 9.3 Acetic anhydn'de 3.1

then poured into 125 cc. of a mixture of water and ice and extracted bymethylene chloride. The extract was washed by a normal solution ofhydrochloric acid, then with water, dried over sodium sulfate, treatedwith animal charcoal, filtered and evaporated to dryness under vacuum. Ayellow resin was obtained which was taken up in 10 cc. of isopropylether.

The solution was cooled then for a period of one hour at a temperaturebetween about and +5 C. The crystals obtained were vacuum filtered,triturated with isopropyl ether and dried at 40 C. 0.776 gm. of ZO-bis-(acetoxymethyl)M-pregnene-ill-dione were obtained. The product was alsopurified by chromatography on 18 silica gel with elution by methylenechloride containing 8% acetone, then recrystallization from isopropylether. The pure product had a melting point of 132 C. and a specificrotation [a] =|-134.4 (c.=0.5% in chloroform).

- The product was soluble in alcohol, acetone, benzene, chloroform and'isopropyl ether (in the hot), insoluble in water and dilute aqueousacids and alkalis.

Analysis.C H O molecular Weight:458.57. Caloulated: C, 70.71%; H, 8.35%.Found: C, 70.6%; H, 8.5%.

This compound is not described in the literature.

STEP C: PREPARATION OF ZO-BIS-(HYDROXY- METHYL) -A*-PREGNENE-3,11-DIONEI'750 mg. of the compound prepared in Step B were introduced into amixture of 37.5 cc. of ethanol and 2.25 cc. of water. The mixture wasagitated under nitrogen and 0.75 cc. of 30% sodium hydroxide solutionwere added. The solution obtained was agitated for a period of three andone-half hours, under nitrogen, at room temperature. The reactionmixture was then poured into 300 cc. of ice water and agitated for aperiod of thirty minutes at +5 C. The precipitate was vacuum filtered,washed with water until the wash water was neutral and dried at -100 C.466 mg. of raw 20-bis-(hydroxymethyl)-A -pregnene-3,11-dione wereobtained which was purified by successive recrystallizations fromethanol.

[Faking into account the recovery of purified product starting from therecrystallization mother liquors, a total yield of the order of 53% wasobtained. The product had a melting point of 208 C. and a specificrotation +166 (c.=0.5% in chloroform).

The product was soluble in ethanol, acetone and chloroform, insoluble inwater, ether and benzene.

Analysis.C 'H O molecular weight=374.5. Calculated: C, 73.76%; H, 9.15%;O, 17.09%. Found: C, 74.1%; H, 9.3%; O, 17.2%.

This compound is not described in the literature.

STEP D: PREPARATION 013 2OBIS-(NITROMETHYL)-A PREGNENE-3,11-DIONE 0.5cc. of 48 B. nitric acid was introduced into 2 cc. of acetic anhydrideunder agitation and under nitrogen with a temperature between -15 and 20C. Then 138 mg. of the compound prepared in Step C in 2 cc. ofchloroform were added. The reaction mixture was maintained underagitation and under nitrogen for a period of twenty minutes at -10 C.The reaction mixture was thereafter poured into 25 cc. of ice water andthe dinitrate was extracted with methylene chloride.

The organic phase was washed in turn with water, with a 2% sodiumbicarbonate solution, then with water, until the wash water was neutral.The organic phase was then dried over sodium sulfate and the solventdistilled to dryness. A clear yellow resin was obtained which set up bythe addition of methanol or of ethanol. The raw product was taken upwith methylene chloride and chromatographed on silica gel with elutionby methylene chloride containing 2% acetone. The resulting solution wastreated with animal charcoal and evaporated to dryness. The residue wasredissolved at reflux in ethanol, concentrated under vacuum,crystallized at room tern perature and vacuum filtered. The precipitatewas washed with iced ethanol and dried at 40 C. s

mg. of 20-bis-(nitnatometh-yl)-A -pregnene-3,11- dionewere obtained,being a yield of 65%. The cornpound had a melting point of 144-146" C.and a specific rotation [a] +l04i2.5 (c.=0.5% in chloroform).

Ultraviolet spectra: (e=l5,300

The product was soluble in acetone, benzene and chloroform, slightlysoluble in ethanol, insoluble in water and ether.

r max. (ethanol)=237 my.

Analysis. C H O N molecular weight=464.5. Calculated: C, 59.46%; H,6.94%. Found: C, 59.6%; H, 7.0%

This compound is not described in the literature.

Example VlH.Preparation of 20-Bis-(Nitrat0methyl)- APregnadiene-3,Il-Dione STEP A: PREPARTION OF 2,4DIBROMO-20 BIS-(ACET-OXYMETHYL) 5,BPREGNANE-3,11-DIONE 2 gm. of20-bis-(acetoxymethyl)-5,8-pregnane-3,11-dione were introduced into 20cc. of acetic acid. Then under agitation and at a temperature betweenabout 20 and 22 C. about 0.2 cc. of acetyl chloride were added andimmediately thereafter, over a period of about ten minutes, 10 cc. of asolution of bromine in acetic acid (testing I l-14.2% bromine) wereadded. The reaction mixture was then poured into 300 cc. of a mixture ofwater and ice.

The crystals obtained were vacuum filtered, washed with water and dried.2.649 gm. of 2,4dibromo-20-bis- (acetoxymethyl)-5fl-pregnane-3,ll-dionewere obtained having a bromine content of 26.6% (theoretical 25 89% Thiscompound was insoluble in water.

This compound is not described in the literature.

The starting compound was prepared by esterifying with acetic acid thehydroxymethylenic functions of 20- bis (hydroxymethyl) -5B-pregnane-3,1l-dione.

STEP B PREPARATION OF 20-BIS-(ACETOXYMETHYL)- A -PREGNADIENE-3,11DIONE1.1 gm. of lithium bromide were introduced into 18 cc. ofdimethylformamide and the mixture was heated under agitation whilebubbling nitrogen therethrough to 80 C. 1.1 gm. of lithium carbonatewere added and thereafter 2.640 gm. of the compound prepared in Step Awere added.

The reaction mixture was heated then to 95-100 C. under agitation andnitrogen for a period of about seventeen to eighteen hours. The reactionmixture was cooled to 20-25" C. and poured slowly into 180 cc. of amixture of water and ice and 3.5 cc. of acetic acid. The precipitate wasvacuum filtered, washed with water and dried.

The raw product was purified by trituration in boiling isopropyl ether,then by chromatography on silica gel with elutions with methylenechloride containing 6% and 8% acetone. The eluates were evaporated andthe residue taken up by hot isopropyl ether and allowed to remainovernight at room temperature. The crystals obtained were vacuumfiltered, washed with isopropyl ether and dried at 80 C.

0.935 gm. of 20-bis-(acetoxymethyl)A -pregnadiene 3,11-dione wereobtained, having a block melting point of 141142 C. and an ultravioletextinction of 15,200 at 239' mm.

The product was soluble in chloroform, slightly soluble in ether andisopropyl ether and insoluble in water.

This compound is not described in the literature.

STEP C PREPARATION OF 2O-BIS-( HYDROXYM'ETHYL)- A -PREGNADIENE-3J1-DIONE0.90 gm. of the compound prepared in Step B were introduced into 22 cc.of 95% ethanol and 2.7 cc. of Water. The solution was maintained for aperiod of fifteen minutes at 2022 C. while bubbling nitrogentherethrough. Then 0.9 cc. of sodium hydroxide solution were introduced.The reaction mixture was maintained for a period of three hours at about20-22 C. under nitrogen. It was then neutralized by addition of aceticacid and poured into 300 cc. of a mixture of water and ice. Theprecipitate formed was vacuum filtered, washed with water and dried.

Purification of the raw product was made by dissolving the product in arefluxing mixture of ethyl acetate and ethanol, filtering the hotsolution and recrystallizing at a temperature between about and C. 0.44gmof 20 bis-(hydroxymethyl)-A -pregnadiene-3,1l-dione, having a meltingpoint of 202-203 C., were obtained.

This compound was soluble in chloroform, slightly soluble in ether andethyl acetate, insoluble in water.

It is not described in the literature.

STEP D: PREPARATION OF 20BIS-(NITRATOMETHYL)- A -PREGNADIENE 3,ll-DIONE1.52 cc. of turning nitric acid were introduced slowly into 6.2 cc. ofacetic anhydride, cooled to a temperature of between 15 and -20 C. Overa period of two to three minutes 0.420 gm. of the compound prepared inStep C in solution in 6.2 cc. of chloroform were added. The reactionmixture was maintained about twenty minutes under agitation and nitrogenbubbled therethrough at l0 to 15 C. and was then poured slowly into cc.of a mixture of water and ice.

The precipitate formed was separated and the liquid was extractedseveral times with methylene chloride. The organic phases were combined,washed successively with water, with a 2% sodium bicarbonate solution,and with water until the wash water was neutral and then dried overmagnesium sulfate.

The solvent was distilled under vacuum and the residue was poured into25 cc. of water and ice and allowed to remain one hour under agitation.The 20-bis-(nitratomethyl)-A -pregnadiene-3,l1-dione crystallized andthe crystals were vacuum filtered, washed with water and dried at 20 C.

The raw product was redissolved in refluxing ethanol and the solutionwas filtered while hot and allowed to crystallize in an ice box.

A new purification was efiected in the same fashion by treatment withanimal charcoal.

0.24 gm. of the pure product were obtained, having a melting point of152-153 C. and a specific rotation [u] =+106:3 (c.=0.5% in chloroform).

This compound was soluble in chloroform, soluble in the hot in ethanol,slightly soluble in ether, insoluble in water.

AnaZysis.C H N O molecular weight=462.49. Calculated: C, 59.72%; H,6.53%; N, 6.05%. Found: C, 59.9%; H, 6.7%; N, 6.3%.

Ultraviolet spectra: A max. (ethanol) 238 m (6: 14,900).

This compound is not described in the literature.

Example IX.Preparati0n of 3a-Ph0sphat0-20-Bis-(Hydroxymethyl)-5-Pregncme-1 1 -One STEP A: PREPARATION OF THE ACETONIDE or 3a-DI-BENYLPHOSPHATO 20 BIS-(HYDROXYMETHYL) -5fi PREGNANE-11-ONE 1.7 g. of theacetonide of 20-bis-(hydroxymethyl)-5flpregnane-3a-ol-11-one, preparedin Step A of Example II, were dissolved in 17 cc. of pyridine. Themixture was cooled to -25 C. and 10 cc. of a solution of dibenzylchlorophosphonate in ether were added. The mixture was agitated at 25 C.under an atmosphere of nitrogen for a period of about 1 hour and heldovernight at -5 C. Next the suspension obtained was poured into cc. of amixture of ice and water. The aqueous suspension was agitated for aperiod of about 1 hour. Hydrochloric acid was added thereto to adjustthe pH to a value of 4, Thereafter, the mixture was extracted withether. The extracts were washed with water, dried over magnesium sulfateand evaporated to dryness. Raw acetonide of 3 ct-dibenzylphosphato 20bis-(hydroxymethy1)-5 ,B-pregnane-l l-one was obtained which was used assuch for the next step of the synthesis.

This compound is not described in the literature.

20BIS- (HYDROXYMETHYL) -5fl-PREGNANE11-ONE1 STEP B: PREPARATION OF3a-DIBENZYLPHOSPHATO- The raw compound from Step A was taken up in amixture of 20 cc. of ethanol and 1 cc. of 6 N hydrochloric acid. Themixture was agitated for about 2 hours and thereafter 200 cc. of waterwere added. The mixture was extracted with ethyl acetate. The extractswere washed with water, dried over magnesium sulfate and evaporated todryness. Raw 3a-dibenzylphosphato ZO-bis-(hydroxymethyl) -5f3-pregnane-ll-one Was obtained which was purified by successive chromatography overmagnesium silicate with elutions by a mixture of methylene chloride andmethanol. 702 mg. of the product were obtained.

. This compound is not described in the literature.

STEP PREPARATION or 3a-PHOSPHATO-20-BIS- (HYDROXYMETHYL)-5fiPREGNANE-ll-ONE1 506 mg. of the compound prepared in Step B wereintroduced into 50 cc. of methanol. 100 mg. of palladized carbon blackcontaining of palladium were added and hydrogen was introduced for aperiod of about 6 minutes, Next the mixture was filtered and thefiltrate was evaporated to dryness under vacuum. Raw 3a-phosphato 20bis-(hydroxymethy1)-5 fl-pregnane-l l-one was thus obtained and wasmoderately soluble in water and insoluble in chloroform.

This compound is not described in the literature.

The said compound was easily transformed into its disodium salt by theaction of sodium methanolate with a methanolic solution of the rawcompound obtained as above to obtain the disodium salt of3a-phosphato-20-bis- (hydroxymethyl) -5}3-pregnane-1 l-one.

This salt is very soluble in water and dilute aqueous alkalis, slightlysoluble in dilute aqueous acids and methanol, insoluble in ether.

Analysis. C I-I O- PNa H O; molecular weight =520.50 (solvated product).Calculated: C, 53.07%; H, 7.55%; P, 5.95%. Found: C, 53.2%; H, 7.7%; P,6.2%.

This compound is not described in the literature. STEP D: PREPARATION OF3a-PHOSPHATO-20-BIS- (NITRATOMETEYL) -513-PREGNANE-1l-ONE 0.4 cc. offuming nitric acid were slowly introduced into 1.2 cc. of acetic acidanhydride cooled to a temperature between -10 C. and C. Then at 10 C.,114 mg. of 30a phosphato -bis-(hydroxymethyl)-5fi-pregnanell-one wereadded. The reaction mixture was held for a period of a half hour between10 C. and -5 C. Then it was poured into 20 cc. of a mixture of water andice and allowed to stand for a period of 1 hour. The precipitate formedwas then separated, washed with water, vacuum filtered and dried undervacuum. 116 mg. of 3aphosphato 2O bis(nitratomethyl)-5fi-pregnaue-1l-one were thus obtained having a meltingpoint of 200 to 220 C. (with decomposition).

The compound Was soluble in chloroform and dilute alkalis, moderatelysoluble in alcohol, very slightly soluble in water and dilute aqueousacids.

This compound is not described in the literature.

The 20 bis (hydroxymethyD-l l-oxygenated-SB-pregnanes and theirderivatives can be used by oral methods in the form of tablets or byparenteral methods in intramuscular injection in the form of aqueous oroily suspensions or even by venous methods in the form of a solution inan adequate excipient and by rectal methods.

They can be made in the form of injectable solutions, injectablesuspension, put up in ampules or in flasks, in tablets and insuppositories.

Preparation 0 Suitable Pharmaceutical Compositions Tablets Containing0.5 Mg.

In an appropriate mixer, there was introduced successively the activecompound, white sugar, potato starch and lactose. On the homogenousmixture there was poured an aqueous solution of gelatin in a quantitysufficient to obtain a mixture capable of being granulated through aperforated metal die.

The granules obtained were placed on a perforated bed on a dish and putin a ventilated dryer at a temperature of 50 C. The dry granules wereground and passed through a metal screen of appropriate dimension, thenmixed with talc and magnesium stearate for lubrication.

The granulated powder was transformed into tablets of adequate weight bymechanical pressure in a press. The tolerance of mean weight of a lot often tablets was :5 per 100 of the theoretical weight. For ingestablecommodities, it was found preferable that the weight of the tablets benot more than 750 mg. nor less than mg. The percentage of activeprincipal in the tablets was found preferably to be approximatelybetween 0.6 and 6 per 1,000.

Pharmacologk al Study of the ZO-Bis-(Hydrdxymethyh-11-OxygenatedJfl-Pregnanes and Derivatives Thereof ACTION ON THECORONARY BLOOD FLOW Study of the action of the said compounds on thecoronary blood flow was eifected on the isolated heart of a rabbit,utilizing a technique inspired from Langendorff (Arch. gesarn. PhysioL,1895, 61, 291). In this method, the heart was suspended by the aorta toa tube and the coronary system was perfused by means of this tube undera constant pressure of 5 om. of mercury by a Locke serum having a pH of7.2 to 7.3, heated to 37 C.

The compound being studied was placed in solution in ethanol and thissolution was diluted by means of the Locke serum to the concentrationusable. By a proper apparatus, the coronary blood flow was registeredand parallelly the ventriculary contractions. The table below summarizesthe results obtained by these compounds, as well as the results fromtrinitrine, papaverine, and other compounds under the same experimentalconditions.

Efieet on the Increase Ventriculary Threshold of Ooro- ContractionsSubstance Active connary Duration N o. centration Blood in mins.

in /cc. Flow in Ampli- Fre- Percent tude in quency in Percent Percent 0.2 20 45 -0 0 0. 05-0. 1 10-15 8-15 -0 -l0 0. 005-0. 01 40-50 30-60 +1010 0. l 30 20 0 l6 0.05 20 20 +20 5 0. 05 10 12 +10 10 005-01 10 10 -0 00. 1 22 15 0 11 0.01 70 40 +25 20 0. 001-0. 005 30 15 0 0 O. 005 10-1515 0 0 0. 05-0. 1 10 s 10 0 1. 0 60 20 30 +15 1. 0 130 23 10 5 1. 0 0 40. 5 25 10 15 0 1.0 20 18 -10 l0 1.0 10 2-20 0 -5 10. 0 20 15-20 0 0 1Slight augmentation.

I. 20-bis-(hydroxymethyD-5fl pregnane-iia-ol-ll-one.

II. Ba-acetoxy-ZO-bis-(hydroxymethyl)-5B-pregnane-11-onc.

III. ZO-bis-(nitratomethyl)-5fl-pregnane-3a-o1-1l-one.

V. 3a-nitrato-20-bis-(nitratomethyl)-5B-pregnane-l1-one.

VI. ZO-bis-(nitratomethyl)-fifi-pregnane-3,1l-dione.

VII. ZO-bis-(acetoxyrnethyl)-A -pregnene-3,11-dione.

VIII. ZO-bis-(hydroxymethyl)-A -pregnene-3,11-dione.

IX. 20 bis-(nitratomethyl)-A"-pregnene-3,11-dione.

X. ZO-bis-(nitratomethyl)-A -pregnadicne-3,11-dione.

XII Potassium salt of 3a-sulfato-20-bis-(nitratornethyl)-5B-pregnonc--one.

XII. ZO-bis-(acetoxymethyl)-5fl-pregnane-3,11-dione.

XIII. Hydrobromide oi papaverine.

XIV. Persantin.

XV. Tetranitrate of pentaerythritol.

XVI. Methyl-3-chromone.

XVII. Khellin.

XVIII. Trinitrine.

XIX. Papaverine.

ACUTE TOXICITY Toxicity tests were made on mice of the Roclcland strainweighing between 18 and 22 gm. The test compounds were used insuspensions containing 10 'mgfper co. in a dispersing solution. Theywere injected in this form by subcutaneous method in groups of ten micein doses of 50, 100 and 200 rug/kg. respectively.

The animals were heldunder observation for one week.

No symptoms of intoxication and mortality were noted in the course ofthis period.

Compound studied: Devoid of toxicity, mg./kg.

III 200 IV 100 V 100 VI 100 VI H 100' X 100 XI 50 XII 100 While thecompounds of the invention are useful in the treatment of angina of thechest and of the coronaritis, they also possess a peripheralvasodilatory action and antispasmodic activity and are therefore usefulin treating asthma, bronchial spasms and arterial spasms.

The present application is a continuation-in-part application ofapplication Serial No. 37,568, filed June 21, 1960, now abandoned, andapplication Serial No. 88,283,

led February 10, 1961, now abandoned.

Various modifications of the process of the products of the inventionmay be made without departing from the spirit or scope thereof, and itis to be understood that the invention be limited only as defined in theappended claims.

C\ CHgOAc OJ 01-13 omoiic wherein R is selected from the groupconsisting of =0 and CH OAG and R; is selected from the group consistingof :0 and and AC is selected from the group consisting of hydro gen,acyl radical of an organic carboxylic acid having 1 to 18 carbon atomsand an anion of a mineral acid.

2. ZO-bis-(hydroxymethyl)-5/3-pregnane-3o-ol-1 l-one. 3.20-bis-(nitratomethy1) -5,8-pregnane-3ot-ol-l l-one.

4. 3oz acetoxy 20 bis (nitratornethyl) 55 pregnane-l l-one.

5. 20 bis (hydroxyrnethyl) A pregnadiene-3,11- dione.

6. 2O bis (acetoxymethyl) A pregnadiene 3,11- dione.

7. 20 bis (nitratomethyl) A pregnadiene 3,11- dione.

8. 20 bis (hydroxyinethyl) 5,8 pregnane 30:,115- diol.

9. ZO-bis-(hydroxymethyl)-5fl-pregnane-3,1l-dione.

10. ZO-bis-(nitratomethyl) -5B-pregnane-3,1 l-dione.

1 1. ZO-bis- (nitratornethyl)-A -pregnene-3,1 l-dione.

12. 20-bis (acetoxymethyl)-A -pregnene-3,1 l-dione.

13. ZO-bis- (hydroxymethyl)-A -pregnene-3,1 l-dione.

14. 3oz nitrato 20 bis (nitratornethyl) 5 8 pregnane-l l-one.

15. 30c sulfato 20 bis (nitratomethyl) 5;? pregnane-l l-one.

16. The alkali metals salts of 3a-sulfato-ZO-bis-(nitratomethyl-5p-pregnane-1 l-one.

17. The potassium salt of 3ot-sulfato-20-bis-(nitratomethyl)-5,6-pregnane- 1 l-one.

18. The pyridine salt of3a-sulfato-ZO-bis-(nitratomethyl)-5,8-pregnane-1l-one.

19. ZO-bis-(acetoxymethyl)-5/3-pregnane-3,1l-dione.

20. 30c acetoxy 20 bis (hydroxymethyl) 5,8-pregnane-l l-one.

21. 30c phosphate 20 bis (hydroxymethyl) 5,8- pregnane-l l-one.

22. 30c phosphato 20 bis (nitratomethyl) 5,8- pregnane-l l-one.

23. The disodium salt of 3a-phosphato-20-bis-(hydroxymethyl-5B-pregnane-1 l-one.

24. A compound having the formula on; CHzOAc II 2 Br wherein R isselected from the group consisting of =0 and H and Ac is selected fromthe group consisting of hydrogen, acyl radical of an organic carboxylicacid having 1 to 18 carbon atoms an anion of a mineral acid.

25. A compound having the formula CH3 01120 R;

wherein R is selected from the group consisting of =0, and

OAc

R and R are selected from the group consisting of hydrogen, phenyl,phenyl-snbstituted lower alkyl and lower alkyl radicals and Ac isselected from the group consisting 25 of hydrogen, acyl radical of anorganic carboxylic 'acid having 1 to 18 carbon atoms and an anion of amineral acid.

26. A compound having the formula CH3 CHzOAO wherein R is selected fromthe group consisting of and CH OAc wherein Ac is selected from the groupconsisting of hydrogen, acyl radical of an organic carboxylic acidhaving 1 to 18 carbon atoms and an anion of a mineral acid whichcomprises condensing 3a-acetoxy-20-formyl-5B- pregnane-l-l-one withformaldehyde to form 20-bis-(hydroxymethyl) -5,8-pregnane-3 OL-Ol-l1-one, reacting said compound with a member of the group consisting ofO=CHR and to form a compound having the formula CH3 CHzO R3 wherein Rand R are selected from the groups consisting of hydrogen, phenyl,phenyl substituted lower alkyl and lower alkyl, oxidizing said compoundto form a compound having the formula wherein R and R have the abovedefinitions, hydrolyzing the latter under acidic conditions to form20-bis-(hydroxymethyl)-5p-pregnane,3,1l-dione and recovering the desiredproducts.

29. A process for the preparation of a compound having the formula omOHzOAc /O\ CH2OA0 I wherein Ac is selected from the group consisting ofhy drogen, acyl radical of an organic carboxylic acid having 1 to 18carbon atoms and an anion of a mineral acid which comprises reacting3a-acetoxy-ZO-formyl-5 8-pregnane-l l-one with formaldehyde to formZO-bis-(hydroxymethyl)-5,8-pregnane-3a-ol1l-one, reacting said productwith a member selected from the group consisting of to form a compoundhaving the formula CH3 CHzO R:

wherein R and R are selected from the groups consisting of hydrogen,phenyl, phenyl-substituted lower alkyl and lower alkyl, reacting thelatter with an acylating agent to form a compound having the formula CH301120 R;

flj onto R4 wherein R is an acyl radical of an organic carboxylic acidhaving 1 to 18 carbon atoms and R and R have the above definitions,hydrolyzing the latter under acidic conditions to form3a-acyloxy-20-bis-(hydroxymethyl)-5,8 pregnane-ll-one, reacting thelatter with an acylating agent to form3a-acyloxy-20-bis-(acyloxymethyl)-5B- pregnane-l1-one, saponifying thelatter to form ZO-bis- (acyloxymethyl)-5B-pregnane 3a-ol-11-one andrecovering the desired compound.

CH3 OHzOAc wherein R is selected from the group consisting of 0 and andAc is selected from the group consisting of hydrogen, acyl radical of anorganic carboxylic acid having 1 to 18 carbon atoms and an anion of amineral acid which comprises reacting ZO-bis-(hydroxymethyl)-11-oxygenated-5,8-pregnane-3-one with an acylating agent to form20-bis-(acyloxymethyl)-1l-oxygenated-Sdpregnane-3-one, brominating thelatter to form 4-bromo-20- bis (acyloxymethyl) ll oxygenated 5,8-pregnane- 3-one, dehydrobrominating the said compound to form 20 bis(acyloxymethyl) ll oxygenated A pregnene-3-one, saponifying the latterto form ZO-bis-(hydroxymethyl)-1l-oxygenated-A -pregnane-3-one andrecovering the desired compound.

31. A process for the preparation of a compound having the formula CH3CHzOAc wherein R is selected from the group consisting of =0 and A0 isselected from the group consisting of hydrogen, acyl radical of anorganic carboxylic acid having 1 to 18 carbon atoms and an anion of amineral acid which comprises brominating20-bis-(acyloxymethyl)-1l-oxygenated- 5,8-pregnane-3-one to form2,4-dibromo-20 bis-(acyloxymethyl) 11 oxygenated 5/3 pregnane 3 one,dehydrobrominating the latter to form 20-bis-(acyloxymethyl)-11-oxygenated-A -pregnadiene-3-one, saponifying the latter to form20-bis-(hydroxymethyl)-ll-oxygenated- A -pregnadiene-3-one andrecovering the desired product.

32. The process of claim 29 wherein a compound having the formula:

wherein R and R are selected from the groups consisting of hydrogen,phenyl, phenyl-substituted lower alkyl and lower alkyl, reacting thelatter with an acylating agent to form a compound having the formula:

C H3 C1120 R3 STATES PATENT OFFICE CERTIFICATE OF CORRECTION PatentNo,-3 l23,597 March 3, 1964 Daniel Bertin et' al, I

It is hereby certified that error appears in the-above. numbered patentrequiring correction and that the said Letters Patent should read ascorrected below.

Column 9, the extreme bottom left-hand formula in the upper right-handportion of the formula and column 10, the} extreme top right-handformula, in the upper right-hand portion of the formula reading \CHBOH HI read \CH2OH column 10, the extreme bottom right-hand formula in theupper right-hand portion of the formula reading:

column 14 line 21, for "pyridene" read pyridine line 63 for."dicarbonate'v read bicarbonate vcolumn 15, line 5. for read 11116 9,f0! "This starting" read The starting column 20, lines 70 and 71, strikeout "20BIS(HYDROXYMETHYL) -5B -PREGNANE- ll-ONE STEP B: PREPARATION OF3q-DIBENZYLPHOSPHATO and insert instead STEP B: PREPARATION OF3q-DIBENZYLPHOSPHATO- 20-BIS(HYDROXYMETHYL) 5{3 PREGNANE-ll-ONE column24-, line 55, after "atoms" insert and Signed and sealed this 5th day ofJanuary 1965.

(SEAL) Attest:

ERNEST w, SWIDER EDWARD J, BRENNER Attesting Officer Commissioner ofPatents

1. A COMPOUND SELECTED FROM THE GROUP CONSISTING OF
 18. THE PYRIDINESALT OF 3A-SULFATO-20-BIS-(NITRATOMETHYL)-5B-PREGNANE-11-ONE.